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The leakage detection system of vials based on two-line water-vapor tunable diode laser absorption spectroscopy (TDLAS) developed in our previous work was first employed to investigate the instantaneous leakage processes of sealed vials. It showed that the leakage states of the vials with medicines cannot be accurately determined in a short time after the opening. Therefore, an optimized system was developed with two-line oxygen TDLAS method by the measurement of defined oxygen leakage coefficient, and it was then utilized to study the